Abstract To develop a sensitive fluorescence polarization immunoassay (FPIA) for detecting Diisobutyl phthalate (DIBP) in Yoghurt. Five tracers with different lengths of bridge between the hapten molecule and fluorescein isothiocyanate (FITC), fluoresceinthiocarbamyl ethylenediamine (EDF), fluoresceinthiocarbamyl hexylenediamine (HDF), were synthesized and their respective binding responses with anti-DIBP-specific antibody were evaluated. The physicochemical parameters were optimised for the FPIA. The FITC-labelled DIBP conjugate (DIBP-FITC) was found to be the optimal tracer for FPIA of Diisobutyl phthalate. The detection limit of DIBP, IC50 value and the working range were 0.82, 9.32 and 1.16–74.97 ng/mL, respectively; and the reaction time was only 5min. The average recovery from spiked Yoghurt samples was 77.80–115.60% which showed good reproducibility for FPIA. Thus, the developed FPIA method exhibited the potential for the rapid and accurate determination of DIBP in agricultural and environmental samples.

中文翻译：

---

基于新型单克隆抗体的荧光偏振免疫分析检测酸奶中的邻苯二甲酸二异丁酯

摘要 开发一种灵敏的荧光偏振免疫分析法（FPIA），用于检测酸奶中邻苯二甲酸二异丁酯（DIBP）。合成了在半抗原分子和异硫氰酸荧光素 (FITC)、荧光素硫代氨基甲酰乙二胺 (EDF)、荧光素硫代氨基甲酰己二胺 (HDF) 之间具有不同桥长度的五种示踪剂，并评估了它们各自与抗 DIBP 特异性抗体的结合反应。物理化学参数针对 FPIA 进行了优化。发现 FITC 标记的 DIBP 偶联物 (DIBP-FITC) 是邻苯二甲酸二异丁酯 FPIA 的最佳示踪剂。DIBP的检出限、IC50值和工作范围分别为0.82、9.32和1.16-74.97 ng/mL；反应时间仅为5分钟。加标酸奶样品的平均回收率为 77.80–115。60% 表明 FPIA 具有良好的重现性。因此，开发的 FPIA 方法显示出快速准确测定农业和环境样品中 DIBP 的潜力。